(A) Time lapse images showing a gradual fragmentation and loss of E-cad network at 73, 80, 85, and 92 hrs-pp; scale bar represents 100 µm (B) (left) The three different cell populations in the aggregate are marked at 72 hrs-pp: namely cells that express only T-Bra, cells that express both T-Bra and E-cad, and cells that express only E-cad. (right) the fraction of the total population of cells for the three classes are provided in the histogram (n = 7 aggregates: 126, 116, 121, 102, 85, 184, and 163 cells, N = 2 replicates) with the SD shown by the error bars; inset shows the spatial distribution of the cells in one such aggregate (right) Spatial connectivity graph is generated for the aggregate at 72 hrs-pp. (C) Spatial distribution of the population fraction of the different cell types as a function of the radial distance from the aggregate centre (n = 7 aggregates: 126, 116, 121, 102, 85, 184, and 163 cells, N = 2 replicates). (D) Islands of E-cad/Oct4 expressing cells surrounded by T-Bra+ cells are present within the aggregate fixed at 92 hr-pp; the inset zooms on a few such islands. (E) Pixel intensity correlation plot between Oct4 and T-Bra (n = 6 aggregates, N = 2 replicates).